[1]索森森,李爱琴,原玲玲,等.巨噬细胞缺氧/复氧及M2型极化对小鼠心肌细胞凋亡的影响及机制[J].新乡医学院学报,2022,39(7):612-616.[doi:10.7683/xxyxyxb.2022.07.003]
　SUO Sensen,LI Aiqin,YUAN Lingling,et al.Effect and mechanism of macrophage hypoxia/reoxygenation and M2 type polarization on cardiomyocyte apoptosis of mice[J].Journal of Xinxiang Medical University,2022,39(7):612-616.[doi:10.7683/xxyxyxb.2022.07.003]
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巨噬细胞缺氧/复氧及M2型极化对小鼠心肌细胞凋亡的影响及机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
39
期数:
2022年7
页码:
612-616
栏目:
基础研究
出版日期:
2022-07-05

文章信息/Info

Title:
Effect and mechanism of macrophage hypoxia/reoxygenation and M2 type polarization on cardiomyocyte apoptosis of mice
作者:
索森森; 李爱琴; 原玲玲; 李世朋; 毋领娟; 柴文文; 宰娇娇; 郭　鹏
（焦作市人民医院心内科，河南　焦作　454000）
Author(s):
SUO Sensen; LI Aiqin; YUAN Lingling; LI Shipeng; WU Lingjuan; CHAI Wenwen; ZAI Jiaojiao; GUO Peng
(Department of Cardiology,Jiaozuo People′s Hospital,Jiaozuo 454000,Henan Province,China)
关键词:
缺血再灌注; 缺氧/复氧; 巨噬细胞; 极化; 心肌细胞; 凋亡; 白细胞介素-10
Keywords:
ischemia reperfusion; hypoxia/reoxygenation; macrophage; polarization; cardiomyocytes; apoptosis; interleukin-10
分类号:
R541.4
DOI:
10.7683/xxyxyxb.2022.07.003
文献标志码:
A
摘要:
目的　探讨心肌缺血再灌注损伤环境下白细胞介素(IL)-10诱导的巨噬细胞M2表型极化对小鼠心肌细胞凋亡的影响及机制。方法　根据培养条件，将小鼠巨噬细胞分为对照组（A组）、缺氧/复氧组（B组）、IL-10组（C组），将小鼠心肌细胞分为对照组（D组）、缺氧/复氧组（E组）、缺氧/复氧+IL-10组（F组）。A组巨噬细胞培养于常氧（O2体积分数为21%）条件下，B组巨噬细胞培养于缺氧（O2体积分数为3%～5%）条件下，24 h后A组和B组巨噬细胞培养于常氧条件下6 h；C组巨噬细胞培养于常氧条件下，并使用含终质量浓度50 mg·L-1 IL-10新鲜完全培养基培养6 h；收集A组、B组、C组巨噬细胞培养基上清液备用。将A组培养基上清液和新鲜完全培养基等体积混合培养D组心肌细胞，将B组培养基上清液和新鲜完全培养基等体积混合培养E组心肌细胞，将B组培养基上清液和C组培养基上清液等体积混合培养F组心肌细胞。采用实时荧光定量聚合酶链式反应法检测A组、B组、C组巨噬细胞中诱导性一氧化氮合酶（iNOS）、CD86、精氨酸酶-1（Arg-1）、CD206 mRNA相对表达量；采用酶联免疫吸附试验检测A组、B组、C组以及B组和C组等体积混合上清液（B+C组）中IL-1β、肿瘤坏死因子(TNF)-α、IL-10、转化生长因子 (TGF)-β蛋白相对表达量；采用流式细胞术检测D组、E组、F组心肌细胞凋亡情况。结果　B组巨噬细胞中iNOS、CD86 mRNA 相对表达量显著高于A组，Arg-1、CD206 mRNA相对表达量显著低于A组（P<0.05）；C组巨噬细胞中Arg-1、CD206 mRNA 相对表达量显著高于A组（P<0.05）；A组与C组巨噬细胞中iNOS、CD86 mRNA相对表达量比较差异无统计学意义（P>0.05）。B组巨噬细胞的培养基上清液中IL-1β、TNF-α蛋白相对表达量显著高于A组，IL-10、TGF-β蛋白相对表达量显著低于A组（P<0.05）；C组巨噬细胞的培养基上清液中TGF-β蛋白相对表达量显著高于A组（P<0.05）；B+C组巨噬细胞的培养基上清液中IL-1β、TNF-α蛋白相对表达量显著低于B组，IL-10、TGF-β蛋白相对表达量显著高于B组（P<0.05）；A组与C组巨噬细胞的培养基上清液中IL-1β、TNF-α蛋白相对表达量比较差异无统计学意义（P>0.05）。D组、E组、F组心肌细胞培养24 h时的细胞凋亡率分别为（18.76±1.11）%、（31.72±1.03）%、（20.51±1.02）%；E组心肌细胞凋亡率显著高于D组（P<0.05），F组心肌细胞凋亡率显著低于E组（P<0.05）。结论　IL-10可通过诱导巨噬细胞M2型极化拮抗缺氧/复氧导致的炎症反应，从而抑制心肌细胞的凋亡。
Abstract:
Objective　To investigate the effect and mechanism of interleukin (IL)-10-induced macrophage M2 phenotype polarization in the environment of myocardial ischemia reperfusion injury on cardiomyocyte apoptosis of mice.Methods　According to the culture conditions,the mouse macrophages were divided into the control group (group A),hypoxia/reoxygenation group (group B),IL-10 group (group C)；the mouse cardiomyocytes were divided into the control group (group D)，hypoxia/reoxygenation group (group E),hypoxia/reoxygenation + IL-10 group (group F).Macrophages in the group A were cultured under normoxia (volume fraction of 21% O2),and macrophages in the group B were cultured in hypoxia (volume fraction of 3%-5% O2)，after 24 hours,macrophages in group A and group B were cultured under normoxic condition for 6 hours;the macrophages in the group C were cultured under normoxia condition for 6 hours,and were cultured in fresh complete medium with a final concentration of 50 mg·L-1 IL-10 for 6 hours;the supernatant of macrophage culture medium in the group A,group B and group C was collected.The medium supernatant in the group A and fresh complete medium were mixed in equal volume to culture cardiomyocytes in group D,and medium supernatant in the group B and fresh complete medium were mixed in equal volume to culture cardiomyocytes in group E,and medium supernatant in the group B and medium supernatant in the group C were mixed in equal volume to culture cardiomyocytes in group F.The relative expression levels of inducible nitric oxide synthase (iNOS),CD86,arginase-1 (Arg-1) and CD206 mRNA in macrophages were detected by real-time quantitative polymerase chain reaction in the group A,group B and group C;the relative expression levels of IL-1β,tumor necrosis factor (TNF)-α,IL-10 and transforming growth factor (TGF)-β protein in the culture supernatant of macrophages in group A,group B,group C and equal volumes of mixed supernatants in group B and group C (group B+C) were detected by enzyme-linked immunosorbent assay；the apoptosis of cardiomyocytes in group D,group E and group F was detected by flow cytometry.Results　The relative expression levels of iNOS and CD86 mRNA in macrophages in the group B were significantly higher than those in the group A,and the relative expression levels of Arg-1 and CD206 mRNA were significantly lower than those in the group A (P<0.05);the relative expressions of Arg-1 and CD206 mRNA in macrophages in the group C were significantly higher than those in the group A (P<0.05);there was no significant difference in the relative expression levels of iNOS and CD86 mRNA in macrophages between the group A and group C (P>0.05).The relative expression levels of IL-1β and TNF-β protein in the medium supernatant of macrophages in the group B were significantly higher than those in the group A,and the relative expression levels of IL-10 and TGF-β protein were significantly lower than those in the group A (P<0.05);the relative expression level of TGF-β protein in the medium supernatant of macrophages in the group C was significantly higher than that in the group A (P< 0.05);the relative expression levels of IL-1β and TNF-β protein in the medium supernatant of macrophages in the group B+ C were significantly lower than those in the group B,and the relative expression levels of IL-10 and TGF-β protein were significantly higher than those in the group B (P<0.05);there was no significant difference in the relative expression levels of IL-1β and TNF-β protein in the supernatant of macrophages between the group A and group C (P>0.05).The apoptosis rate of cardiomyocytes cultured for 24 hours in the group D,group E and group F was (18.76±1.11)%,(31.72±1.03)%and (20.51±1.02)% respectively;the apoptosis rate of cardiomyocytes in the group E was significantly higher than that in the group D (P< 0.05);the apoptosis rate of cardiomyocytes in the group F was significantly lower than that in the group E (P< 0.05).Conclusion　IL-10 can antagonize the inflammatory response induced by hypoxia/reoxygenation by inducing M2 type polarization of macrophages,thereby inhibiting cardiomyocyte apoptosis.

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更新日期/Last Update: 2022-07-05